Purification of exosome-like vesicles from urine.

Methods in enzymology ·Vol. 524 ·2013-00-00 ·Pages 225-41

Chen CY, Hogan MC, Ward CJ

Abstract

Urinary exosome-like vesicles (ELVs), 20-200nm membrane-bound particles shed by renal epithelium, have been shown to interact with the primary cilia of distant epithelial cells of the nephron. These ELVs are emerging as an important source of protein, mRNA, and miRNA biomarkers to monitor renal disease. However, purification of ELVs is compromised by the presence of large amounts of the urinary protein Tamm-Horsfall Protein (THP). THP molecules oligomerize into long, double-helical strands several microns long. These linear assemblies form a three-dimensional gel which traps and sequesters ELVs in any centrifugation-based protocol. Here, we present a purification protocol that separates ELVs from THP and divides urinary ELVs into three distinct populations.

MeSH Terms

Biomarkers/chemistry Centrifugation, Density Gradient Electrophoresis, Polyacrylamide Gel Exosomes/chemistry Humans Protease Inhibitors/chemistry Sucrose Urinary Tract/chemistry,metabolism Urine/chemistry Uromodulin/isolation & purification Urothelium/chemistry,metabolism

Chemicals

Biomarkers Protease Inhibitors UMOD protein, human Uromodulin Sucrose

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Chen Christopher Y

Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, USA.

Hogan Marie C

Ward Christopher J

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Article Info

Journal

Methods in enzymology

Abbr.

Methods Enzymol

ISSN

1557-7988

DOI

10.1016/B978-0-12-397945-2.00013-5

Published

2013-00-00

Pages

225-41

Language

English

Region

United States

NLM ID

0212271

PMCID

PMC4028690

Subset

Grants

NIDDK NIH HHS · P30 DK090728 · United States

NIDDK NIH HHS · R01 DK080688 · United States

External Links

PubMed source DOI: 10.1016/B978-0-12-397945-2.00013-5 PMC Full Text

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